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Expressed truncated N-terminal variable surface glycoprotein (VSG) of Trypanosoma evansi in E. coli exhibits immuno-reactivity

机译:在大肠杆菌中表达的伊氏锥虫的截短的N末端可变表面糖蛋白(VSG)表现出免疫反应性

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摘要

The variant surface glycoprotein (VSG) of trypanosome is an important part of its body surface coat, which is expressed in early, middle and late stages of infection contributing a major diagnostic value. In the present study, the 5' end of the partial VSG gene sequences (681 bp) encoding N-terminal protein of RoTat 1.2 VSG (227 amino acid) was amplified, cloned into pET32a vector, and expressed in prokaryotic system. The fused His-tagged expressed VSG protein (43 kDa) of the Trypanosoma evansi was characterized in SDS-PAGE and immunoblotting using hyperimmune/immune sera raised against buffalo, dog, lion and leopard isolates of T. evansi. The expressed protein remained immunoreactive with all the sera combinations. The animals immunized with whole cell lysate or recombinant protein showed similar antibody reactions in ELISA and CATT (Card Agglutination Test for Trypanosomiasis). This study suggests the expressed recombinant truncated VSG is having its importance for its possible use in sero-diagnosis of surra. (C) 2012 Elsevier B.V. All rights reserved.
机译:锥虫体的表面糖蛋白变异体(VSG)是其体表层的重要组成部分,在感染的早期,中期和晚期表达,对诊断具有重要价值。在本研究中,扩增了编码RoTat 1.2 VSG(227个氨基酸)N端蛋白的VSG部分基因序列(681 bp)的5'端,并将其克隆到pET32a载体中,并在原核系统中表达。埃文锥虫的融合有His标签的表达的VSG蛋白(43 kDa)在SDS-PAGE和免疫印迹中进行了鉴定,使用针对伊文氏牛的水牛,狗,狮子和豹分离株的超免疫/免疫血清。表达的蛋白质对所有血清组合均保持免疫反应性。用全细胞裂解物或重组蛋白免疫的动物在ELISA和CATT(锥虫病卡片凝集试验)中显示出相似的抗体反应。这项研究表明,表达的重组截短的VSG对于其在Surra的血清学诊断中的可能用途具有重要的意义。 (C)2012 Elsevier B.V.保留所有权利。

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